title: Fluctuations in butyrate-producing bacteria in ulcerative colitis patients of North India.
abstract: OBJECTIVE
  To study the interplay between butyrate concentration and butyrate-producing bacteria in fecal samples of ulcerative colitis (UC) patients vs control individuals.
  
  
  METHODS
  Fecal samples were collected from 14 control individuals (hemorrhoid patients only) and 26 UC patients (severe: n = 12, moderate: n = 6, remission: n = 8), recruited by the gastroenterologist at the Department of Gastroenterology, All India Institute of Medical Sciences, New Delhi, India. Disease activity in UC patients was determined by clinical colitis activity index. We employed fluorescent in situ hybridization in combination with flow cytometry to enumerate the clostridium cluster population targeted by 16S rRNA gene probe. Major butyrate-producing species within this cluster were quantified to see if any change existed in control vs UC patients with different disease activity. This observed change was further validated by quantitative polymerase chain reaction. In addition to this, we carried out gas chromatography to evaluate the changes in concentration of major short chain fatty acids (SCFAs), namely acetate, n-butyrate, iso-butyrate, in the above samples. Student t test and Graph pad prism-6 were used to compare the data statistically.
  
  
  RESULTS
  There was a significant decrease of Clostridium coccoides (control, 25.69% ± 1.62% vs severe, 9.8% ± 2.4%, P = 0.0001) and Clostridium leptum clusters (control, 13.74% ± 1.05% vs severe, 6.2% ± 1.8%, P = 0.0001) in fecal samples of UC patients. Furthermore, we demonstrated that some butyrate-producing members of the clostridial cluster, like Fecalibacterium prausnitzii (control, 11.66% ± 1.55% vs severe, 6.01% ± 1.6%, P = 0.0001) and Roseburia intestinalis (control, 14.48% ± 1.52% vs severe, 9% ± 1.83%, P = 0.02) were differentially present in patients with different disease activity. In addition, we also demonstrated decreased concentrations of fecal SCFAs, especially of n-butyrate (control, 24.32 ± 1.86 mmol/μL vs severe, 12.74 ± 2.75 mmol/μL, P = 0.003), iso-butyrate (control, 1.70 ± 0.41 mmol/μL vs severe, 0.68 ± 0.24 mmol/μL, P = 0.0441) and acetate (control, 39.51 ± 1.76 mmol/μL vs severe, 32.12 ± 2.95 mmol/μL, P = 0.047), in the fecal samples of UC patients. The observed decrease of predominant butyrate producers of clostridial clusters correlated with the reduced SCFA levels in active UC patients. This was further confirmed by the restoration in the population of some butyrate producers with simultaneous increase in the level of SCFA in remission samples.
  
  
  CONCLUSIONS
  Our observations indicate that decreases in members of the clostridial cluster resulting in reduced butyrate levels contribute to the etiology of UC.
authors.affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India.
authors.forename: Reena
authors.initials: R
authors.lastname: Kumari
authors.forename: Vineet
authors.initials: V
authors.lastname: Ahuja
authors.forename: Jaishree
authors.initials: J
authors.lastname: Paul
mesh_terms: D000085:Acetates / Q000378:metabolism; D000328:Adult; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D058491:Bacterial Load; D002087:Butyrates / Q000378:metabolism*; D016022:Case-Control Studies; D002849:Chromatography, Gas; D003013:Clostridium / Q000254:growth & development / Q000378:metabolism; D003093:Colitis, Ulcerative / Q000175:diagnosis / Q000382:microbiology*; D003106:Colon / Q000382:microbiology*; D005243:Feces / Q000382:microbiology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007194:India; D058610:Isobutyrates / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D021521:Ribotyping
chemical_list: D000085:Acetates; D002087:Butyrates; D058610:Isobutyrates
keywords: Clostridial cluster; Fecal microbiota; Fluorescent in situ hybridization-flow cytometry; Quantitative polymerase chain reaction; Short chain fatty acids; Ulcerative colitis